Effect of Nicotinic Acid on Sperm Characteristic and Oocyte Development after In Vitro Fertilization using Cryopreserved Boar Semen

초록

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilizationability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezingsolution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawedsuch as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PIdouble staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double stainingfor mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDAstaining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in5 and 10 mM (61.1 ± 1.5%, 64.7 ± 2.0%) of nicotinic acid than other groups (0 mM, 52.1 ± 2.3%; 20 mM, 47.8± 5.1%, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid(26.1 ± 1.8%, 24.9 ± 1.5%) than other groups (0 mM, 35.3 ± 0.8%; 20 mM, 36.5 ± 1.9%, P<0.05). The live spermwith mitochondrial integrity was significantly higher in 5 and 10 mM (84.2 ± 3.6%, 88.4 ± 2.3%) of nicotinic acidthan other groups (0 mM, 77.3 ± 4.4%; 20 mM, 73.3 ± 3.6%, P<0.05). Blastocyst rate of in vitro development wassignificantly higher in 10 mM (17.0 ± 1.3%) of nicotinic acid than other groups (0 mM, 9.4 ± 0.5%; 5mM, 12.6 ±0.8%; 20 mM, 5.0 ± 1.0%, P<0.05). Moreover, total cell number was higher in 5 and 10 mM (53.6 ± 2.9%, 57.9± 2.8%) of nicotinic acid than other groups (0 mM, 41.0 ± 1.4%; 20 mM, 23.2 ± 2.8%, P<0.05). Hydrogen peroxidein embryos was lower in 5 mM nicotinic acid (0.7 ± 0.1%) than other groups (0 mM, 1.0 ± 0.1%; 10mM, 0.9 ±0.0%; 20 mM, 1.4 ± 1.0%, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and qualityof spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation

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